Designing of the expressing eukaryotic plasmid of the G1 epitope of bovine ephemeral fever virus G glycoprotein in human embryonic kidney cells

The G glycoprotein of bovine ephemeral fever virus (BEFV) has been identified as a plausible candidate for immunization against BEF disease. In recent years, this protein has been investigated to produce a recombinant vaccine. The aim of the present study was to construct a eukaryotic plasmid, expressing G1 epitope of BEFV G glycoprotein gene, for application as a possible DNA vaccine in future studies. For this purpose, the G1 epitope of G glycoprotein gene was cloned in a eukaryotic expression vector, pEGFP-N1, under the control of the human cytomegalovirus (CMV) promoter. Then, the recombinant pEGFPN1-G1 construct was transfected into human embryonic kidney (HEK 293) cell line and the expression efficiency was verified by indirect immunofluorescent (IFA) staining and immunoblotting techniques. Observation of intracytoplasmic fluorescence in transfected cells and appearance of a distinct band at an approximate molecular weight of 26 kDa in immunoblotting in reaction to an anti-G1 mouse serum, indicated that G1 protein was successfully expressed by this recombinant construct in the host cells.